Asparagine deamidation in
peptides or in
fibronectin fragments containing the
asparagine-glycine-arginine sequence generates
isoaspartate-
glycine-
arginine (isoDGR), a new
alphavbeta3 integrin-binding motif. Because alphavbeta3 is expressed in angiogenic vessels, we hypothesized that isoDGR-containing
peptides could be exploited as
ligands for targeted delivery of drugs to
tumor neovasculature. We found that a cyclic CisoDGRC
peptide coupled to fluorescent nanoparticles (
quantum dots) could bind
alphavbeta3 integrin and colocalize with anti-CD31, anti-alphavbeta3, and anti-alpha5beta1
antibodies in human
renal cell carcinoma tissue sections, indicating that this
peptide could efficiently recognize endothelial cells of angiogenic vessels. Using CisoDGRC fused to
tumor necrosis factor alpha (TNF) we observed that ultralow doses (1-10 pg) of this product (called isoDGR-TNF), but not of TNF or CDGRC-TNF fusion
protein, were sufficient to induce antitumor effects when administered alone or in combination with
chemotherapy to
tumor-bearing mice. The antitumor activity of isoDGR-TNF was efficiently inhibited by coadministration with an excess of free CisoDGRC, as expected for
ligand-directed targeting mechanisms. These results suggest that isoDGR is a novel
tumor vasculature-targeting motif.
Peptides containing isoDGR could be exploited as
ligands for targeted delivery of drugs, imaging agents, or other compounds to
tumor vasculature.